Metabolic profiling of single cells by exploiting NADH and FAD fluorescence via flow cytometry

Abir AH, Weckwerth L, Wilhelm A, Thomas J, Reichardt C, Munoz LE, Völkl S, Appelt U, Mroz M, Niesner R, Hauser A, Fischer RS, Pracht K, Jäck HM, Schett G, Krönke G, Mielenz D (2024)

Publication Language: English

Publication Type: Journal article, Original article

Publication year: 2024

Journal

Molecular Metabolism Elsevier

Book Volume: 87

Article Number: 101981

DOI: 10.1016/j.molmet.2024.101981

Abstract

Objective: The metabolism of different cells within the same microenvironment can differ and dictate physiological or pathological adaptions. Current single-cell analysis methods of metabolism are not label-free. Methods: The study introduces a label-free, live-cell analysis method assessing endogenous fluorescence of NAD(P)H and FAD in surface-stained cells by flow cytometry. Results: OxPhos inhibition, mitochondrial uncoupling, glucose exposure, genetic inactivation of glucose uptake and mitochondrial respiration alter the optical redox ratios of FAD and NAD(P)H as measured by flow cytometry. Those alterations correlate strongly with measurements obtained by extracellular flux analysis. Consequently, metabolically distinct live B-cell populations can be resolved, showing that human memory B-cells from peripheral blood exhibit a higher glycolytic flexibility than naïve B cells. Moreover, the comparison of blood-derived B- and T-lymphocytes from healthy donors and rheumatoid arthritis patients unleashes rheumatoid arthritis-associated metabolic traits in human naïve and memory B-lymphocytes. Conclusions: Taken together, these data show that the optical redox ratio can depict metabolic differences in distinct cell populations by flow cytometry.

Authors with CRIS profile

Ariful Haque Abir Department of Molecular Immunology Leonie Weckwerth Department of Molecular Immunology Artur Wilhelm Department of Medicine 3 – Rheumatology and Immunology Jana Thomas Department of Molecular Immunology Clara Reichardt Department of Medicine 3 – Rheumatology and Immunology Luis Munoz Becerra Department of Medicine 3 – Rheumatology and Immunology Simon Völkl Department of Medicine 5 – Haematology and Oncology Rebecca Sophie Fischer Department of Molecular Immunology Katharina Pracht Department of Molecular Immunology Hans-Martin Jäck Professur für Immunologie Georg Schett Lehrstuhl für Innere Medizin III Gerhard Krönke Department of Medicine 3 – Rheumatology and Immunology Dirk Mielenz Lehrstuhl für Innere Medizin III

Involved external institutions

Deutsches Rheuma-Forschungszentrum (DRFZ) DE Germany (DE) Charité - Universitätsmedizin Berlin DE Germany (DE)

How to cite

APA:

Abir, A.H., Weckwerth, L., Wilhelm, A., Thomas, J., Reichardt, C., Munoz, L.E.,... Mielenz, D. (2024). Metabolic profiling of single cells by exploiting NADH and FAD fluorescence via flow cytometry. Molecular Metabolism, 87. https://doi.org/10.1016/j.molmet.2024.101981

MLA:

Abir, Ariful Haque, et al. "Metabolic profiling of single cells by exploiting NADH and FAD fluorescence via flow cytometry." Molecular Metabolism 87 (2024).

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